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Isolation and Characterization of Coumarin Isolated from Endophyte, Alternaria Species -1 of Crotalaria pallida and Its Apoptotic Action on HeLa Cancer Cell Line

Abstract

Umashankar T, Govindappa M, Yarappa Lakshmikantha R, Padmalatha Rai S and Channabasava

Endophyte, Alternaria species-1 isolated from leaf part of Crotalaria pallida was selected in isolation, identification and purification of coumarin. HPLC, UV-spectrophotometry, FTIR, NMR and XRD studies were carried out on characterization of endophytic fungal coumarin. In vitro apoptotic activity was carried out against HeLa cervical cancer cell lines. The results of MTT studies, acridine staining activity revealed the coumarin as an effective in inducing apoptotic activity in HeLa cells. The coumarin significantly inhibited the proliferation of HeLa cells and it’s a concentration and time dependent manner. Significant elevation of caspase 3 and 9 and activity was observed in coumarin treated HeLa cells but no elevation or activity of caspase 7, 8, 10 was not observed. Coumarin has shown 0.156 μg/ml of significant IC50 value against viable of HeLa cell lines. Thus, coumarin exerts apoptotic activity by caspase dependent apoptosis by enhancing the caspase 3 and 9 and it degraded the DNA (avoided the further replication). This is the first report in internationally that coumarin isolated from endophyte, Alternaria species-1, purified and characterized and its role was identified in apoptotic activity.